Sirisha & Sujathamma RJLBPCS 2018 www.rjlbpcs.com Life Science Informatics Publications Original Review Article DOI: 10.26479/2018.0405.27 IN VITRO PROPAGATION OF FEW PIMPINELLA SPECIES: A REVIEW K. Bala Sirisha, P. Sujathamma Department of Biosciences and Sericulture Sri Padmavathi Mahila Visvavidyalayam (Women’s University) Tirupati, India. ABSTRACT: The genus Pimpinella L. is one of the largest genera in Apiaceae subfamily Apioideae, with approximately 150 species distributed in Asia, Europe, and Africa. A few species can also be found in South America and one can be found in the western part of North America. The present study includes four species of Pimpinella (Pimpinella anisum, P. tirupatiensis, P. candolleana and P. brachycarpa). Maximum of Pimpinella species can be annual, biennial or perennial. They are generally characterized by the presence of fibrous collars on the top of the rootstock and compound umbels. These plants usually grow on dry rocky places, rocky crevices, fields, meadows, mountain pastures, and grasslands. Majority species have more medicinal and Pharmacological properties, generally propagated through seeds. The seed germination in nature is very poor. So, it is difficult for propagating them rapidly to meet the market demand. Hence they are endangered and slowly becoming extinct. Therefore, plant tissue culture techniques may facilitate their propagation over a shorter period of time than conventional techniques. This may also be of value to the preservation and conservation of the genetic resources of this important family. KEYWORDS: Apiaceae, Conservation, Micropropogation, Pimpinella species. Corresponding Author: Dr. K. Bala Sirisha* Ph.D. Department of Biosciences and Sericulture Sri Padmavathi Mahila Visvavidyalayam (Women’s University) Tirupati, India. Email Address: [email protected] 1.INTRODUCTION The genus Pimpinella L., consists of 150 species, distributed throughout the world [1] is one of the largest genera of the family Apiaceae. About two dozen Pimpinella species occur in India. Out of these, eight species are largely confined to peninsular India, eleven occur in the Eastern Himalaya © 2018 Life Science Informatics Publication All rights reserved Peer review under responsibility of Life Science Informatics Publications 2018 Sept - Oct RJLBPCS 4(5) Page No.336 Sirisha & Sujathamma RJLBPCS 2018 www.rjlbpcs.com Life Science Informatics Publications and two are endemic to the western Himalaya [2]. These are annual, biannual, and perennial species usually growing on dry rocky crevices, fields, meadows, mountain pastures and grasslands[3]. Pimpinella belongs to the family Apiaceae and represents many important medicinally valued plants. These are used for pharmaceutical, cosmetics, perfumery and food industry. Some of the species of Pimpinella contains essential oil used as antispasmodic, antioxidant, antimicrobial, insecticidal and antifungal effects[4] . Classification of Pimpinella L. Kingdom : Plantae Subkingdom : Tracheobionta Super division : Spermatophyta Division : Magnoliophyta Class : Magnoliopsida Sub class : Rosidae Order : Apiales Family : Apiaceae Genus : Pimpinella We checked scientific studies in various electronic databases (Medline, Pubmed, Science Direct, Scopus, Google Scholar websites, Books and Various Thesis) from 1993 to 2017. After a comprehensive search on the micropropagation aspects of Apiaceae family in India, we reviewed available publications that provided information about different applications of these plant species in human and livestock. In this article, scientific and author names of plant species were confirmed for latest changes according to “The plant list” (http://www.theplantlist.org)[25]. Morphology, Distribution and Micropropagation studies of Pimpinella species Pimpinella anisum L. Pimpinella anisum L is an aromatic, annual grassy herb with 30-50 cm height, white flowers and small green to yellow seeds with secondary feather like leaflets of bright green, twice pinnate[5],[6]. Distribution P. anisum present in Eastern Mediterranean region, West Asia, the middle East, Mexico, Egypt and Spain. Micropropagation “Somatic embryogenesis” from callus cultures has been reported [7]. A detailed description of P. anisum embryogenesis through seed culture has been reported [8]. The cell culture of P.anise was grown in the presence or absence of (2,4-D) [9]. Application of isopentenyladenine or isopentenyladenosine (4.10-8 to 4.10-7 M) to the proembryonic culture yielded an increase of the cell density, in contrast to a proembryonic culture grown without exogenous application of cytokinins. Embryogenesis was induced by transferring the cells to a hormone-free medium. Embryo © 2018 Life Science Informatics Publication All rights reserved Peer review under responsibility of Life Science Informatics Publications 2018 Sept - Oct RJLBPCS 4(5) Page No.337 Sirisha & Sujathamma RJLBPCS 2018 www.rjlbpcs.com Life Science Informatics Publications development was promoted by isopentenyladenine and isopentenyladenosine (5.10-8 to 5.10-7 M), higher concentrations (5.10-6 M) inhibited embryogenesis. The effect of cytokinins on embryogenesis was only promotive until the third day of culture, i.e. coincident with cell growth rather than differentiation. In vitro development of P.anise clonal lines, root explants cultured on Murashige and Skoog’s media supplemented with 4 different growth regulators was reported [10]. The synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) induced somatic embryos from callus. Napthalenacetic acid (NAA), also a synthetic auxin, induced prolific root formation. The synthetic cytokinins 6-Benzylaminopurine (BAP) and Thidiazuron (TDZ) induced adventitious shoot formation. The 2,4-D induced somatic, embryogenesis system followed by plant regeneration was the best system for the establishment of clonal lines of P.anise. Simple in vitro protocol for large scale multiplication of plants from various explants of P. anisum has been developed [11]. High frequency of multiple shoots formation was achieved from callus cultures derived from shoot apices, root, and stem explants, and also from seed derived calli. Somatic embryogenesis was observed in callus cultures derived from seed and shoot apices. Complete plants developed from these embryoids. Direct regeneration of plantlets from shoot apices was also observed. Root formation occurred in all the cultures. The requirement for exogenous auxins and cytokinin for differentiation was found to be varying in different tissues. It was observed that hairy root cultures in P.anisum L. using cultures were grown in four different media, both under darkness and under photo period conditions of 16h light [12]. A high frequency organogenesis in anise (P.anisum L.) using hypocotyl explants from in vitro germinated seedlings was reported [13]. Both cytokinins, benzylaminopurine and kinetin induced shoot regeneration but the effect of Benzylaminopurine (BAP) was more pronounced. High frequency shoot regeneration (45 shoots explant-1) was obtained on BAP 1 mg l1. Both cytokinins were also tested in combination with auxins (Naphthalene acetic acid /Indole acetic acid). Interaction of benzyl amino purine /kinetin with naphthalene acetic acid/ indoleacetic acid increased the length of regenerated shoots. Different kinds of callus morphology were observed but it had no relationship with regeneration potential. The regenerated plants were normal and healthy. In vitro micropropagtion of P.anisum L. by using various explants shoot tip, node, internode and seeds was done [14]. It was reported that mature seeds of P.anisum cultured on Murashige and Skoog’s Basal medium (MSBM) fortified with GA3 (2.89 µM/L) was suitable for seed germination (77.3±1.0%). In shoot tip culture, shoot tip measuring 1-2 cm was excised from 20 days in vitro axenic plants of P.anisum and observed that cultured on MSBM with BAP (13.3µM) and NAA (5.37 µM) was the best medium for shoot tip initiation and formation of multiple shoots, MSBM with BAP (13.3µM) and NAA (5.37 µM) was the best medium for initiation and formation of multiple shoots from nodal explants, MSBM with 2,4,-D (2.26 µM) was the best medium for induction and growth of whitish green callus from the internodes as explants of P.anisum, when compared to the callus derived from MSBM with IAA and MSBM with NAA in different © 2018 Life Science Informatics Publication All rights reserved Peer review under responsibility of Life Science Informatics Publications 2018 Sept - Oct RJLBPCS 4(5) Page No.338 Sirisha & Sujathamma RJLBPCS 2018 www.rjlbpcs.com Life Science Informatics Publications concentrations. (Table: 1) Pimpinella tirupatiensi Bal. & Sub. Pimpinella tirupatiensis is locally known as ‘adavikothimeera’ (Forest Coriander). It is a narrow endemic species of seasonal occurrence with underground tuberous root system [15]. It is an erect herb with perennial tuberous root stock, stem is simple, branched, terete, striate; branches alternate, bifurcate; branch lets glabrous, veins prominent, margins cartilaginously crenate - serrate. Basal leaves are simple, ovate obtuse or acute, deeply cordate, 1.7-3.8 x 1.3-3.8 cm, petiole 2.5-7.8 cm long, sheathing at base, cauline leaves palmately 3-partite . Flowers are white, 5-16, in compound umbles bracteoles 1-2, very small, linear. Petals are 1 mm long, glabrous, obovate, sub- orbicular, apex inflexed; styles small, slender, 1mm long; stylopod conical, yellowish brown, persistent. Fruits are 1.5 mm long, ovoid, Papillos - scrabrous [16].